Genetic variation of somaclonal mutants from the 8th generation of Pekalongan accession rodent tuber (typhonium flagelliforme lodd.) based on RAPD-PCR analysis

Abstract

Rodent tuber is a potential plant which can be developed as an anticancer drug. The rodent tuber plant has a low anticancer compound which does not have an economic value to be utilized on an industrial scale, particularly to explore as an anticancer drug. The material plant was used 30 clones of Rodent Tuber Pekalongan to increase its bioactive compounds using a combination of in vitro culture with gamma-ray irradiation. The selection process has been done until the 8th generation and obtained 14 somaclonal mutants that have higher bioactive compounds than control plant (non-mutant). This study aimed to analyze the genetic variation of 14 somaclonal mutants of rodent tuber using RAPD with 14 primers. The results were produced 513 total bands with the size of 200-5000 bp and 380 polymorphic bands between somaclonal mutants. OPB-18 primer produced the most polymorphic bands and five specific polymorphic bands of 1336 bp, 1070 bp, 901 bp, 861 bp and 728 bp in the somaclonal mutant M23 (20-4-2-1-1-1). OPD-20 primer produced seven specific polymorphic bands of 1574 bp, 1557 bp, 1501 bp, 1496 bp, 1234 bp, 1229 bp and 1086 bp in the M23 somaclonal mutant. The phylogenetic analysis showed that there were two main groups with the coefficient similarity between 0.77 and 0.83. The highest genetic variation obtained was found in the M23 somaclonal mutant with a genetic difference of 23% compared with control. There are five somaclonal mutants (M24, M22, M21, M16, M14) that have a genetic variation of up to 22% over the control. This study shows that OPB-18 and OPD-20 primers were efficient in detecting somaclonal mutant variants.